Then sera from immunized mice were diluted before added into the wells and incubated for 2 h at 37 °C. Plates were then washed with washing buffer (PBS-0.05% Tween 20) three times for 3 min each and goat anti-mouse IgG was added into the wells and incubated for 1 h at 37 °C. After washing as above, TMB (3, 3′,5,5′-tetramethylbenzidine dihydrochloride) substrate

(Sigma) was added and color intensity was determined spectrophotometrically at OD 450 nm. Statistical analysis was performed by Gehan-Breslow-Wilcoxon Test using Graphpad Prism 5. P≤ 0.05 was regarded as significant. 19 proteins associated with S. aureus invasion or pathogenesis were dotted onto NC membranes and reacted with sera from mice recovered from infection with different S. aureus strains. The sera from Caspase inhibitor mice infected with S. aureus 1884 reacted with proteins FnBA, SasA, SasF and SPA (S. aureus proteins A) (Fig. 1A). The sera from mice infected with S. aureus 546 reacted with proteins

CNA, FnBA, SasA, SasF, and SPA(Fig. 1B). The sera from mice infected with S. aureus USA300 reacted with proteins ClfA, IsdA, SasA and SPA (Fig. 1C). We found different S. aureus strains induced different antibody responses. The proteins SasA and SPA reacted with all of the sera. Protein SPA is a mitogen that interacts with many immunoglobulins by binding to the Fc region. The results showed that SasA was expressed on all of the above strains and could induce strong antibody response during S. aureus infection. LY2157299 datasheet To detect whether the antibody response induced by SasA is protective, part of the protein was expressed. The SasA protein is composed of 2272 amino acids. The secondary structure of SasA protein was analyzed by DNAstar software and fragment (48aa-333aa, named fSasA) was selected to be amplified from the genomic DNA of S. aureus USA300 by PCR using primers SasAF and SasAR. Recombinant plasmid pET-fSasA was constructed, sequencing verified, and transformed into E. coli BL21 for protein expression. After induction with 1 mM IPTG at 37 °C for 4 h, the total soluble proteins of bacteria were analyzed by SDS-PAGE. It showed

that fSasA was expressed at a level of up to 10% of whole cell protein (Fig. 2A). After 2-step purification, fSasA protein of high purity was obtained (Fig. 2B). Western blot with antibody against 6x His tag showed that Lenvatinib price the protein size was correct (Fig 2C). The purified protein can be used as antigen for animal immunization. SasA is a surface protein of S. aureus. The fSasA was absorbed well by aluminium hydroxide gel in physiological saline. After second immunization, BALB/c mice generated strong IgG response against fSasA protein. The response was further elevated after third immunization (Fig. 3). To test the role of immunity induced by fSasA against S. aureus infection, BALB/c mice were challenged with 3 × 109 S. aureus USA300, collected at early exponential phase, 7 days after the third immunization with fSasA.

6B). Thus, the cell surface structures sialoadhesin and B7-H1 are involved in the induction of the IL-35+ Treg. We demonstrate

in this study that IL-35 production and release is induced Opaganib manufacturer in human peripheral blood CD4+ and CD8+ T cells by B7-H1 and sialoadhesin co-stimulation, provided by DC. Such IL-35+ T cells are potent Treg, which, in contrast to IL-10-driven type-1 Treg (Tr1), do not suppress T-cell responses via IL-10 and/or TGF-β 11. Several pieces of evidence support the conclusion that the R-DC-induced Treg act via IL-35. Neutralization with anti-EBI3 and anti-p35 Ab and depletion of IL-35 removed the inhibitory effect of the SN of Treg and naïve T cells from CB, which do not produce IL-35 upon stimulation with R-DC, lack suppressor function. Thus, induction of IL-35 represents a novel activation program in human T cells responding to viral infection. EBI3 is a member of the IL-12 family. It was first identified in B lymphocytes based on its induction following EBV infection. It codes for a 34 kDa-secreted glycoprotein homologous to the p40 subunit of IL-12. Recent studies have shown that EBI3

can dimerize with IL-12 p35 and EBI3/p35 was called IL-35. The in vivo association between EBI3 and p35 was originally evidenced in human placental extracts Tamoxifen 20. Data presented in Fig. 4 and 5 demonstrate that IL-35 and not IL-27 or even IL-12 is responsible for the inhibitory effect of the SN. More recent studies demonstrated that IL-35 is constitutively expressed by mouse CD4+CD25+FOXP3+ Treg 3, 5. Transcripts coding for EBI3 and p35 were observed to be constitutively coexpressed by mouse Treg and EBI3/p35 heterodimer Aldehyde dehydrogenase was coprecipitated from the cell culture SN of these cells. In addition, in vitro and in vivo studies suggested that the expression of IL-35 by mouse Treg contributed to their suppressive function 21. However, human CD4+CD25+FOXP3+ Treg do not constitutively express IL-35 and induction of FOXP3 upregulates neither EBI3 nor p35 mRNA in human T cells 6, 7. Yet, recombinant mouse IL-35 was shown to inhibit

the proliferation of mouse effector T cells in vitro. In another recent study, a single chain mouse IL-35-Fc fusion protein was demonstrated to enhance the proliferation of mouse Treg, while inhibiting the development of Th17 cells 5. The data of this study demonstrate for the first time that IL-35 is a potent regulatory cytokine, also in the human immune system, and that a combinatorial signal delivered from DC to T cells via B7-H1 and sialoadhesin is crucial to the induction of human IL-35+ Treg. We observe transient FOXP3 expression in T cells stimulated by R-DC as well as DC. Such temporal activation-induced FOXP3 expression in human T cells has been described before and is not obligatory correlated with a regulatory function, whereas natural CD4+CD25+ Treg show constitutive FOXP3 expression 10, 22.

The baseline characteristics of the patients were similar in the two groups (Table 1). The number of episodes of moderate-massive haemoptysis during the study period did not

differ significantly between the groups (four in each group). The total number of radiological interventions (two bronchial artery embolisation procedures in each group) and the number of surgical procedures (three in itraconazole group and four in the control group) were also similar in the two groups during the trial. The number of patients showing overall response was higher in the itraconazole group (76.5%) compared with the control group (35.7%), and was statistically significant (P = 0.02). The learn more numbers of patients demonstrating a clinical response and radiological response (Fig. 2) were also significantly higher Romidepsin mouse in the itraconazole group (Table 2). The mean

longest diameter of pulmonary lesions in the itraconazole and control groups, respectively, was 32.4 (13.9) and 28.2 (11.7) mm, and 26.3 (9.1) and 32.4 (9.7) mm at baseline and 6 months respectively. Adverse events were noted in 8 (47.1%) patients in the itraconazole group, however, none was serious and none led to any discontinuation of the study drug. Transient abnormality of liver function was noted in two patients in the itraconazole group. In both the cases, the liver enzymes were elevated between 1.5 and 2 times the upper limit of normal. The liver function was found to be deranged at the second and third month of therapy, respectively, in the two patients. The liver functions normalised Protirelin on follow-up in these two patients despite continuation of itraconazole therapy. Gastrointestinal disturbances were documented in six patients in the itraconazole group. All the patients were followed up for a median (IQR) of 11 (7–16) months after completion of the trial. On follow-up, 9/17 (5 of 13 with overall response) and 10/14 (1 of 5 with overall response) patients worsened

in the itraconazole and control group respectively. There was radiological and clinical worsening in six and clinical worsening alone in four patients in the control group, whereas there was radiological and clinical worsening in seven and clinical worsening alone in two patients in the itraconazole group. During the follow-up four patients died, two in each group. Two patients died from uncontrolled massive haemoptysis, one patient died from postoperative sepsis whereas one patient died due to acute coronary syndrome. Our initial search retrieved 372 citations, of which 19 studies have evaluated the role of antifungal agents in CPA (Table 3).[2, 10-13, 17-30] The studies have utilised oral (itraconazole, voriconazole, posaconazole) and intravenous (amphotericin B, itraconazole, voriconazole, micafungin) antifungal agents in patients with CPA. The overall response ranges from 14% to 93% with the response lower in patients with CCPA and highest in those with CNPA (Table 4).

The innate immune system contributes to airway inflammation in

asthma [13] and is mediated by activated leucocytes, including eosinophils [14, 15], mast cells [14, 15], CD4+ T lymphocytes [16] and B cells [17]. It is well established that IgE plays a major role in asthma and allergic reactions through its ability to bind to Fc-epsilon receptor I on mast cells [18]. In this study, we demonstrate that GTE and purified EGCG suppress in vitro induction of human IgE responses in a dose-dependent fashion, suggesting a potential and safe therapeutic option for treating asthma selleck products and other diseases of altered IgE regulation. Study participants.  Peripheral blood (40 ml) was obtained from n = 3 allergic asthmatic patients (2 men and 1 woman, age 30–45 years selleck compound old), from the State University of New York (SUNY) Downstate Asthma Center of Excellence (Table 1). Asthmatic patients presented with clinically defined severe-persistent asthma (e.g. have asthma symptoms throughout the day, use rescue inhaler multiple times a day, have a FEV1 <60% of predicted)

[19], atopy (skin prick positive to at least one of the following panel: Ragweed [Short, Tall], 5 Grass Mix [Timothy, Orchard, June, Red Top, Sweet Vernal], English Plantain, 10 Tree Mix [Ash, Beech, Birch, Elm, Hickory, Maple, Oak, Poplar, Sycamore, Alder]) and perennial allergens Dust Mite (Dermatophagoides pteronyssinus and/or Dermatophagoides farinae, Hollister-Stier, Spokane, WA), American Cockroach, Cladosporium, Alternaria, Dog epithelium, and/or cat pelt (Alk-Abello, Round Rock, TX, USA), and allergic rhinoconjunctivitis, with elevated serum IgE levels

(681–2368 IU/ml). None of the subjects received allergen immunotherapy within the prior 6 months. Asthma treatment regimen included as needed inhaled β-agonists and corticosteroids. Written informed consent was obtained from the study participants. The study was approved by the SUNY Downstate Medical Protirelin Center Institutional Review Board, and the procedures followed were in accordance with institutional guidelines involving human subjects. Total Serum IgM, IgG, IgA.  Blood was collected and immunoglobulin (Ig) levels (IgM, IgG, IgA) were detected in serum. All serum Ig determinations were carried out using nephelometry performed according to manufacturer’s recommendations in the Clinical Diagnostic Laboratory at SUNY Downstate Medical Center (reference range for healthy adult serum: IgM: 47–367 mg/dl; IgG: 648–2045 mg/dl; IgA: 55–375 mg/dl; IgE: 20–100 IU/ml.) Total Serum IgE.  Blood was collected and immunoglobulin E (IgE) levels were determined using the UniCap Total IgE fluroenzyme immunoassay (Pharmacia and Upjohn Diagnostics) performed according to the manufacturer’s recommendations (reference range for healthy adult serum: 20–100 IU/ml).

However, new data showed that the Treg-cell pool can remain self-sustained over months 27. Recently, comprehensive high throughput (HT) sequencing studies revealed a very high TCR diversity in human Treg cells, comparable to other T-cell subsets including naïve T cells 28. This led us to the hypothesis that broad TCR diversity may be important for Treg-cell homeostasis and immuno-regulatory function. To address this, we compared highly

diverse Treg cells from WT mice with less diverse Treg cells derived from Rag-sufficient TCR-transgenic (TCR-Tg) mice. In the latter, endogenous TCR rearrangements permit the generation of natural Treg cells with a polyclonal, Smoothened antagonist albeit narrower, TCR repertoire compared with WT mice. Therefore, TCR-Tg mice turned out to be a valuable tool for analyzing the physiological impact of TCR diversity on Treg-cell function. In this system, we performed adoptive transfer experiments and revealed a robust homeostatic advantage of WT Treg cells in TCR-Tg recipients with a less complex Treg-cell repertoire. Such sustained survival and expansion of transferred Treg cells allowed us to recover sufficient numbers of WT Treg cells to correlate their TCR sequences and organ-specific distribution. Furthermore, check details we analyzed the influence of TCR repertoire size on in

vitro suppressive capacity of Treg cells and compared these results with their ability to suppress allogeneic T-cell responses in an in vivo model of lethal acute GvHD. We conclude that, within

the limitations of an IL-2-dependent homeostatic niche, TCR diversity is required for optimal Treg-cell homeostasis and suppressive function. selleck kinase inhibitor In this study, we used Rag-sufficient OT-II TCR transgenic mice in which the TCR repertoire of Treg cells is limited to non-clonotypic ‘escapees’ that are selected on endogenous Tcrb and/or Tcra rearrangements. To monitor and sort Foxp3+ Treg cells, we crossed male homozygous TCR-Tg and female Foxp3-eGFP reporter mice. Male F1-offspring are hemizygous for Foxp3-eGFP and carry the pre-rearranged TCR. GFP+ Treg cells in TCR-Tg mice expressed no or only low levels of the clonotypic TCR and are selected for endogenous TCR rearrangements (Supporting Information Fig. 1) 29, 30. These observations and previous studies of Treg cells with restricted TCR rearrangement options 7, 12, 31 supported the hypothesis that Treg-cell repertoires of TCR-Tg mice are diverse but narrower than those of congenic WT mice. HT sequencing has recently become available to comprehensively characterize TCR repertoires on the level of nucleotide sequences. We chose primers spanning the variable region between the constant Cα and 12 V-elements of the Vα8 (also TRAV12) family.

In the absent reference comprehension literature, there is growing evidence that infants’ ability to locate the absent referent depends on various spatial factors. Some of the factors are the accessibility of the hiding location (Ganea, 2005), its proximity to the infant (Ganea & Saylor, 2013; Saylor & Baldwin, 2004) and, most central

to the present discussion, the stability of object location (Huttenlocher, 1974; Saylor & Ganea, 2007). The current study shows that location information may affect infants’ absent reference comprehension indirectly through affecting their object representation. Encountering an object several times across different locations affects infants’ understanding of the object identity, and this impairs their ability to locate the hidden object upon the experimenter’s verbal request. An interesting question Autophagy inhibitor for

future research is whether this effect can be extended to other types of referents that are less likely to have duplicates, for example to people or objects that infants know are unique. Another question is whether highly salient features that naturally help infants identify objects can release them from the location Palbociclib molecular weight change effect. Finally, it would be interesting to know when in development such type of location change stops interfering with infants’ performance and to understand what cognitive factors lead to such improvement. Previous research has shown that infants are able to individuate objects based on featural information before 12 months, at 4.5–10 months depending on the procedure (McCurry, Wilcox, & Woods, 2009; Wilcox, 1999; Wilcox & Baillargeon, 1998; Wilcox & Woods, 2009; Xu & Carey, 1996). In the current study, 12-month-old infants were confused about the number of objects when not given consistent spatiotemporal information and when their attention was not deliberately drawn to surface features. Several

aspects of the current study design might have contributed to L-NAME HCl this. First, the time lag between the two object presentations was much larger (10 min) in this study than in object individuation studies (a few seconds). Second, infants in this research had not only to individuate an object (establish its representation as a distinct solid entity in space), but also to identify it (that is, bind different object features together that define its identity and hold them in memory throughout occlusion for future retrieval). It is known that object identification is a more challenging task than object individuation (Tremoulet et al., 2000). Third, in the current study, infants’ object recognition was assessed in response to a verbal request for the object when it was absent. Presumably this is a more demanding test situation.

The rats were separated into four groups, each composed of 10 individual rats: (i) 10 mg/kg SLD-treated CLP group; (ii) 20 mg/kg SLD-treated CLP group; (iii) CLP group; and (iv) sham-operated control group. The groups were housed in separate cages. A CLP polymicrobial sepsis model

was applied to the rats, induced through caecal ligation and two-hole puncture. Anaesthesia was induced through the intraperitoneal administration of thiopental 25 mg/kg. The abdomen was shaved and the peritoneum was click here opened. Once the diaphragm exposed the abdominal organs, the caecum was isolated and ligated with a 3/0 silk ligature just distal to the ileocaecal valve. Two punctures were made with a 22-gauge needle through the caecum distal to the point of ligation, and the caecum was returned to the peritoneal cavity. The abdominal incision was then closed with a 4/0 sterile synthetic absorbable suture. The wound was bathed in 1% lidocaine solution to ensure analgesia. The sham-operated group received laparotomies,

and the rats’ caeca were manipulated but not ligated or perforated. All the animals were given 2 ml/100 g body weight of normal saline subcutaneously at the time of surgery and 6 h afterwards for fluid resuscitation. Immediately after the surgical procedure was completed, the rats in the sham-operated and the SLD-treated CLP groups received 10- or 20-mg/kg doses of SLD, which were administered with an oral gavage suspended in saline. There are many sildenafil Epigenetics inhibitor doses for rats, varying from 0·4 mg/kg to 90 mg/kg, with different administration routes [28–33]. The reason we selected 10- and 20-mg/kg doses of oral sildenafil is that 10 mg/kg/day of sildenafil would result approximately

in the same BCKDHA plasma concentration as 50 mg in humans [34]. These doses are very common for rats, and we first aimed to determine if it is protective in CLP-induced organ damage, as well as how the dose affects protection. Therefore, we used 10- and 20-mg/kg oral doses of sildenafil, as have previous authors [35–37]. An equal volume of saline was administered to the sham-operated control group and the CLP group. The rats were deprived of food postoperatively but had free access to water for the next 16 h, until they were killed. The survival rate in CLP-induced sepsis models varies according to the size of the needle used [38]. Otero-Anton et al. reported that mortality after CLP in rats increased gradually with the size of the caecal puncture. They evaluated 0·5-cm blade incision; 13-gauge, 16-gauge and 18-gauge puncture; and four punctures with a 22-gauge needle. Mortality increased gradually with the puncture size, from 27% with a 22-gauge needle to 95% with the blade incision during a week of observation [38]. In addition, in our previous studies we observed mortality within 12–20 h after sepsis induction with a 12-gauge needle [39–42]. However, in studies performed with 21- and 22-gauge needles, mortality was not as common [38,43,44].

In the present experiments, baseline SkBF was lower and the early peak was higher at T2, in comparison with T0, in apparent contradiction with our previous study, where no change in any of these two variables could be detected [3]. The most likely explanation for this apparent discrepancy is the higher number of enrolled subjects (28 vs 12), leading to a greater power to detect relatively small effects.

Desensitization to NO could account for the observed modification of baseline SkBF if, in these thermal conditions (i.e., 34°C), NO actually contributed to lower dermal microvascular tone, as suggested by some [12,16], although not all studies [10,11]. More difficult to understand in this context is the increase in the early peak response observed from T0 to T2. As the early peak SCH772984 is not caused by NO, it should not be affected by removing or attenuating (by desensitization) the action of this mediator. One might argue that the basal level of NO-dependent vasodilation (i.e., in normothermia, prior to heating and during the first few minutes of heating, when it would remain unaffected) Atezolizumab mouse might still modulate the early peak. In that case, however, the expected result of desensitization to NO would be a decrease, not an increase

of the initial vasodilatory response to the thermal stimulus. Some insight into this matter may be provided by data indicating that local heating activates sympathetic nerve endings in the skin microcirculation, with potentially a dual effect on vascular tone, vasodilator on one hand through stimulation of endothelial alpha2 adrenergic receptors

leading to enhanced activity of eNOS, vasoconstrictor on the other hand through a direct action on vascular smooth muscle [8,9]. Importantly, the local thermal challenge seems to dynamically alter the balance between these two effects, tipping it in favor of vasodilation during the first 30 minutes, and in the opposite direction later on, accounting for a progressive decline of SkBF even when local heating is maintained (the “dying out” phenomenon) [8]. We speculate that, in the present study, the first thermal challenge at T0 had a persistent influence on local adrenergic mechanisms, such Arachidonate 15-lipoxygenase that, on the second thermal challenge at T2, the balance was more intensely tipped toward vasodilation at the time of the early peak. Following this line of thought, one might also wonder whether the later tipping of sympathetic influences toward vasoconstriction might not have contributed to lower the plateau response at T2. Clearly, further studies are warranted to test these hypotheses. A final note is required regarding the fact that both the nadir and the plateau responses were somewhat lower when thermal hyperemia was elicited by the commercial, in comparison with the custom-made chamber (Figure 3).

Tissue sections were deparaffinized and pretreated with 0.3% hydrogen peroxide in methanol for 30 min at room temperature to block endogenous peroxidase click here activity. For staining with anti-p62/SQSTM1 antibody, antigens were retrieved by heating sections at 80°C in 10 mmol/L citrate buffer, pH 6.0, for 3 h prior to the hydrogen peroxide treatment. After non-specific binding was blocked with 10% normal goat serum (NGS), sections were incubated with primary antibodies at 4°C overnight.

All antibodies were diluted in 10% NGS. Sections were washed in PBS and then incubated with secondary anti-rabbit or anti-mouse IgG antibodies conjugated to horseradish peroxidase (Envision+ System, DakoCytomation) at room temperature for 1 h. Reactions were visualized with 0.4 mg/mL 3,3′-diaminobenzidine (DAB) in PBS containing 0.006% H2O2 for 10 min. Nuclei were counterstained with hematoxylin. Transmission electron microscopy

was performed as previously described.[7] NVP-BGJ398 mouse Formalin-fixed specimens were dissected into 1 mm3 pieces, and were then post-fixed with 2.5% glutaraldehyde in 0.1 mol/L phosphate buffer (PB; pH 7.4) for 4 h and 1% OsO4 in PB at 4°C for 1 h. Specimens were dehydrated using a graded series of alcohols and QY-1 (Nisshin EM Co., Ltd, Tokyo, Japan), and then embedded in Quetol 812 (Nisshin EM). Ultrathin sections were cut with an LKB ultramicrotome (LKB-Produkter, Bromma, Sweden), and sections were counterstained with aqueous TI-blue (Nisshin EM) and Sato’s lead citrate.[8] Sections were examined using a 1200EX transmission electron microscope (JEOL Ltd, Tokyo, Japan). Written informed consent was obtained from the patient’s parents for the genomic analysis and for publication of the results. Genomic DNA was extracted from frozen liver and spleen using standard protocols. PCR primers were designed to amplify all the exons of NPC1 and flanking intron

regions. Direct sequencing G protein-coupled receptor kinase of PCR products was performed using a 3130xl genetic analyzer (Applied Biosystems, Foster City, CA, USA), and sequence data were analyzed as previously described.[9] At autopsy, the spleen weighed 169 g, slightly heavier than usual. The liver weighed 1058 g and hepatomegaly was not apparent. The pancreas was hemorrhagic in the head, body and tail, indicative of acute hemorrhagic pancreatitis. The brain weight was 731 g. Gross neuropathological findings included marked atrophy of the frontal and temporal lobes bilaterally (Fig. 2a), cerebellum, brainstem and spinal cord. Coronal sections of the cerebrum exhibited marked atrophy of the deep white matter with thinning of the corpus callosum, marked atrophy of the frontal and temporal cortices and mild to moderate atrophy of the parietal and occipital cortices.

011) (Table S2). APS I relatives also had lower number of these cells, although only borderline significant (P = 0.050). Invariant CD1d-restricted NKT-cells (iNKT), Tanespimycin which are supposed to play an inhibitory role in autoimmune diseases, were identified with the help of several characteristic surface markers as Vα24, Vβ11, CD161 and the Invariant NKT-molecule (Table S2). In contrast to Tregs, we did not observe any alterations in iNKT cells in our patients with APS I. Contrary to a previous report [16], the suppressor subset characterized by the markers CD8+CD11b+CD28+ revealed no significant

differences between our studied groups. Further, we analysed several effector/memory T cell subtypes in patients with APS I and their relatives in comparison with control individuals. MS 275 We first confirmed that the percentages of T cells, T helper cells (CD4+) and cytotoxic T cells (CD8+) were similar in patients and controls (Table S2). Unexpectedly, we observed that APS I family members had significantly decreased frequency of memory Th cells (CD4+CD45RA−CD45RO+) compared to healthy controls (P = 0.023) (Table S2, Fig. 2). Next,

we sought to compare the frequency of Th cell subsets with different homing properties according to differentially expressed chemokine receptors. CCR6 and CXCR3 were of particular interest as CCR6+ cells are attracted to epithelial surfaces by CCL20 and can be involved in protection against CMC; CXCR3-expressing cells are attracted to inflammatory tissues by binding to interferon-induced

GPX6 chemokines CXCL9-11 [26, 27]. We did not find alterations in the proportions of CD4+CD45RA−CCR4+CCR6+ lymphocytes that have been reported to contain IL-17A-secreting Th17 cells (Table S2). In contrast, the percentage of CCR6 and CXCR3 coexpressing Th subpopulation, which includes among others IFNγ and IL-17A coproducing cells, was significantly decreased in patients with APS I (P = 0.035) (Fig. 3) [26]. Next, we examined the abundance of myeloid cell subsets in patients with APS I. DC can be subdivided into several undergroups, here separated into MDC1, MDC2 and PDC. PDC differ from MDC in both the expression of pattern recognition receptors, cytokine receptors, cytokines and migration capability [28]. MDC1 are capable of differentiating to Langerhans cells whereas MDC2 cells are not [29]. No differences in the frequencies of dendritic cells were seen in our study (Table S2). Contrary to previous reports, the proportion of monocytes, as determined by CD14 expression in the compartment of live cells purified by Lymphoprep, showed no deviations between patients, controls and relatives. However, large individual variations were seen. When characterizing the monocyte subpopulations, we found that relatives had trends towards less CD14+CD11b+ than their control group (P = 0.053) (Table S2).